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Cytochrome c oxidase levels in chondrocytes during monolayer expansion and after return to three dimensional culture.

Abstract：

OBJECTIVE:Here we investigate whether monolayer culture or culture at 21% oxygen influences activity of cytochrome c oxidase, the terminal enzyme in the respiratory chain whose activity is essential for oxidative metabolism and whether return to three dimensional (3-D) culture restores cytochrome c oxidase activity to original levels. METHODS:Primary bovine articular chondrocytes were cultured in alginate beads (3-D) for 4 weeks or in monolayer under 1% and 21% oxygen for up to 9 days and then returned to 3-D culture for up to 4 weeks. Cells were stained to localise cytochrome c oxidase within the cells. Mitochondrial protein content and cytochrome c oxidase enzymatic activity were determined. Expression of cytochrome c oxidase subunits, COXI and COXIV, was assessed by qRT-PCR. RESULTS:Cytochrome c oxidase staining remained minimal in chondrocytes cultured in alginate for 4 weeks under 21% oxygen. Mitochondrial protein content and cytochrome c oxidase activity increased significantly during 9 days of chondrocyte expansion in monolayer, accompanied by up-regulation of the COXI mitochondrial gene but not the COXIV nuclear-encoded gene. Cytochrome c oxidase staining increased from day 5 of monolayer culture and remained high even after the cells were returned to 3-D culture for 4 weeks. CONCLUSIONS:Culture of chondrocytes in monolayer leads to a rapid increase in mitochondrial protein content and cytochrome c oxidase activity. The increase in cytochrome c oxidase activity is not reversed even after chondrocytes are returned to 3-D culture for 4 weeks; high oxygen tension alone does not appear to stimulate cytochrome c oxidase activity.

journal_name

Osteoarthritis Cartilage

journal_title

Osteoarthritis and cartilage

authors

Boubriak OA,Brooks JT,Urban JP

doi

10.1016/j.joca.2009.03.001

subject

Has Abstract

pub_date

2009-08-01 00:00:00

pages

1084-92

issue

8

eissn

1063-4584

issn

1522-9653

pii

S1063-4584(09)00061-2

journal_volume

17

pub_type

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